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Biomedical subjects

W L Taylor

Publications and source records attributed to W L Taylor.

27 records · Page 2Linked to original sources

Sequence analysis of a cDNA coding for a pancreatic precursor to somatostatin.

A synthetic oligonucleotide having the sequence d(T-T-C-C-A-G-A-A-G-A-A) deduced from the amino acid sequence Phe-Phe-Trp-Lys of somatostatin-14 was used to prime the synthesis of a cDNA from channel catfish (Ictalurus punctatus) pancreatic poly(A)-RNA. The major product of this reaction was a cDNA fragment of 565 nucleotides. Chemical sequence analysis of the cDNA fragment revealed that it was complementary to a mRNA coding for somatostatin. The 565-nucleotide cDNA hybridizes strongly with a poly(A)-RNA estimated to be 1000 nucleotides in length. An amino acid sequence of the somatostatin precursor was predicted from the nucleotide sequence. Oyama et al. [Oyama, H., Bradshaw, R. A., Bates, O. J. & Permutt, A. (1980) J. Biol. Chem. 255, 2251-2254] have reported the isolation of a somatostatin from the catfish that is 22 residues in length (somatostatin-22). This peptide differs from somatostatin-14 in amino acid sequence. The cDNA sequence obtained by this laboratory codes for somatostatin-14 and predicts another somatostatin gene product from this species. Thus it would appear that there are at least two somatostatin gene products.

Animals↗

Purification and characterization of a thyrotropin-releasing hormone deamidase from rat brain.

This report describes the purification of a rat brain thyrotropin-releasing hormone (TRH) deamidating enzyme to apparent homogeneity. Criteria for purity include sodium dodecyl sulfate and disc gel electrophoresis, as well as isoelectric focusing (pI = 4.5). Enzyme purification was facilitated by development of a rapid and sensitive continuous assay using the substrate L-pyroglutamyl-Nim-benzylhistidyl-L-prolyl-beta-naphthylamide, which, upon hydrolysis of the naphthylamide, results in the appearance of the fluorescent product, beta-naphthylamine (beta NA). With this substrate the homogeneous enzyme had a specific activity of 14.5 mumol of beta NA min-1 mg-1. The only peptide product formed was shown to be L-pyroglutamyl-Nim-benzylhistidyl-L-proline. Hydrolysis of [L-prolyl-2,3-3H]TRH was shown to yield L-pyro-glutamyl-L-histidyl-L-proline as the only radiolabeled product. Characterization of the brain deamidase by gel filtration chromatography and sodium dodecyl sulfate gel electrophoresis indicated that the enzyme consists of a single polypeptide chain having molecular weights of 70,000 and 73,500, respectively. Rat brain TRH deamidase has an apparent Km of 34 micron, and a pH optimum between 7 and 8 using L-pyroglutamyl-Nim-benzylhistidyl-L-prolyl-beta-naphthylamide as a substrate. With this substrate, TRH was shown to be a competitive inhibitor with an apparent Ki of 120 +/- 20 micron.

Amidohydrolases↗

The inhibition of thyrotropin-releasing hormone deamidation in porcine hypothalamic tissues.

Frozen porcine hypothalamic fragments afford an enzymic fraction that forms exclusively pyroGlu-His-Pro by enzymatic degradation of thyroliberin. Ten compounds have been examined as inhibitors of this reaction and the results indicate that the enzyme(s) responsible for this conversion have an absolute sulfhydryl requirement. Concentrations of thyroliberin have also recently been reported in areas of the brain in addition to the hypothalamus. Each of these areas actively degrade thyroliberin with the hypothalamus being most active.

Animals↗